Phosphorylcholine-based hydrogel for immobilization of biomolecules. Application to fluorometric microarrays for use in hybridization assays and immunoassays, and nanophotonic biosensing.
Zeneida Díaz-BetancorMaría-José BañulsFrancisco Javier SanzaRafael CasquelMaria Fe LagunaMiguel HolgadoRosa PuchadesÁngel MaquieiraPublished in: Mikrochimica acta (2019)
An approach is presented for covalent immobilization of biomolecules on an acrylate phosphorylcholine hydrogel. The immobilization and the hydrogel formation take place simultaneously by a thiol-acrylate coupling reaction, induced by UV-light (254 nm). The hydrogel is prepared on two polymeric surfaces (the HardCoat protective layer of Blu-Ray discs, and SU-8) and applied to fluorescence microarray and label-free interferometric detection. For the first, Cy5 labeled analytes are used (λem 635 nm) and, for the second, a periodic array of high-aspect ratio nanopillars detects unlabeled analytes by interferometry. Bioavailability of the immobilized probes is demonstrated in labeled assays; for the case of oligonucleotides by discriminating single nucleotide polymorphisms, and, for the case of antibodies, by BSA immunorecognition. The raw hydrogel is employed to detect human C-reactive protein, in both labeled and non-labeled assay formats, with sensitivities of 30 ng·mL-1 and 2 pg·mL-1, respectively. Graphical abstract Schematic presentation of the phosphorylcholine (MPC) hydrogel preparation onto BluRay disc and SU-8 nanopillars to perform fluorescence and label-free interferometric detection, respectively. It selectively detects C-reactive protein (CRP), but it can covalently immobilize antibodies or nucleid acid probes to detect other analytes.
Keyphrases
- label free
- drug delivery
- hyaluronic acid
- wound healing
- tissue engineering
- high throughput
- single molecule
- pet imaging
- small molecule
- photodynamic therapy
- endothelial cells
- cancer therapy
- computed tomography
- fluorescence imaging
- living cells
- escherichia coli
- high resolution
- mass spectrometry
- cystic fibrosis
- high speed
- nucleic acid
- loop mediated isothermal amplification
- electron transfer